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Movement and Structure of Atmospheric Populations of Fusarium

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Movement and Structure of Atmospheric Populations of Fusarium Movement and Structure of Atmospheric Population of Fusarium Binbin Lin Dissertation submitted to the faculty of the Virginia Polytechnic Institute and State University in partial fulfillment of the requirements for the degree of Doctor of Philosophy In Plant Pathology, Physiology, and Weed Science David G. Schmale III, Chair Shane D. Ross, Co-Chair Antonius B. Baudoin Boris A.Vinatzer May 8, 2013 Blacksburg, VA Keywords: Fusarium, plant pathogen, aerobiology, aerobiological sampling, Burkard volumetric sampler, BVS, unmanned aerial vehicle, UAV, inoculum source, long-distance transport Movement and Structure of Atmospheric Population of Fusarium Binbin Lin ABSTRACT Fusarium is one of the most important genera of fungi on earth. Many species of Fusarium are well-suited for atmospheric dispersal, yet little is known about their aerobiology. Previous research has shown that large-scale features known as atmospheric transport barriers (Lagrangian coherent structures) guide the transport and mixing of atmospheric populations of Fusarium. The overall goal of this work is to expand our knowledge on the movement and structure of atmospheric populations of Fusarium. The first objective was to monitor changes in colony forming units (CFUs) in atmospheric populations of Fusarium over small time intervals (10 min to several hours). We hypothesized that consecutive collections of Fusarium with unmanned aerial vehicles (UAVs) demonstrate small variations in colony counts. To test this hypothesis, sampling devices on UAVs were separated into two groups, four inner sampling devices opened during the first 10 minutes and four outer sampling devices opened during the second 10 minutes. Results indicated that (1) consecutive collections of Fusarium at 100 m demonstrated small variations in counts and (2) the similarity between collections decreased as the time between sampling intervals increased. The second objective was to determine the structure of atmospheric populations of Fusarium species and relate this to potential source regions. We hypothesized that diverse atmospheric populations of Fusarium are associated with multiple source regions. To test this hypothesis, Fusarium samples were collected with UAVs and identified to the level of species by sequencing a portion of the translation elongation factor 1- alpha gene (TEF-1). Potential source regions were identified using the atmospheric transport model HYSPLIT. Results indicated that (1) diverse atmospheric populations of Fusarium appeared to be associated with multiple source regions, and (2) the number of Fusarium species collected with UAVs increased with back-trajectory distance of the sampled air. The third objective was to examine the associations between concentrations of populations of Fusarium at ground level (1 m) and in the lower atmosphere (100 m). We hypothesized that concentrations of Fusarium in the atmosphere vary between 1m and 100m. To test this hypothesis, Fusarium was collected with a Burkard volumetric sampler (BVS) and UAVs. Colony counts were converted to spore concentrations (spores per cubic meter of air). Sampling efficiency was used to correct spore concentrations. Results indicated that (1) the distribution of spore concentrations was similar for both samplers over different times of the day, (2) spore concentrations were generally higher in the fall, spring, and summer, and lower in the winter, and (3) spore concentrations were generally higher with BVS samplers than those with UAVs for both hourly and seasonal data. The fourth objective was to assess the ability of strains of Fusarium collected in the lower atmosphere to cause plant disease. We hypothesized that certain isolates of Fusarium collected with UAVs cause plant diseases. To test this hypothesis, we randomly selected isolates of three different species (F. circinatum, F. avenaceum, and F. sporotrichioides) of Fusarium collected with UAVs to inoculate three different hosts (wheat, corn, and pine). Known Fusarium strains were obtained from J. Leslie at Kansas State University as controls. Results indicated showed that the three different isolates tested were able to cause plant diseases in three different hosts (wheat, corn, and pine), confirming that these were potential agents of disease. This work sets the stage for future work examining potential source regions, transport distances, and seasonal patterns of Fusarium. An increased understanding of the dynamics and population structure of plant pathogenic Fusarium in the lower atmosphere is essential for predicting the spread of plant disease and optimizing disease management strategies in the future. iii Acknowledgements It is a great pleasure to thank everyone who has helped me write my dissertation successfully. I would first like to thank my advisor Dr. David Schmale for his guidance, support, and encouragement throughout this research and also with my professional development. I would also like to thank my co-advisor Dr. Shane Ross for numerous scientific discussions, helpful advice, and guidance. My thanks also go to my committee members, Dr. Anton Baudoin and Dr. Boris Vinatzer, for their detailed and constructive comments in the completion of this degree. Also, I would like to thank Dr. John Jelesko for helping to analyze the population structure of Fusarium. I am also thankful to colleagues and friends whose advice, support, and friendship have contributed to the completion of my research. A special thanks to these people include (in no particular order), John Cianchetti, Melissa Keller, Piyum Khatibi, Amir Bozorgmagham, Niki McMaster, Diane Reaver, AJ Prussin, Dashzeveg Gantulga, Hope Gruszewski, Zolton Bair, Zach Upchurch, Brycen Hill, Gunjune Kim, and Megan LeBlanc. I am also thankful to the PPWS staff members including Donna Ford, Patsy Niece, Judy Fielder, and Cris Thompson for all their help throughout this process. A special thanks to Yiming Peng, one of the LISA collaborators in the Department of Statistics, for helping data analysis. Finally, I am forever grateful to my family and friends for their encouragement and support throughout all my studies, which allowed me to continue during the most difficult times in my research and academic obligations. iv Table of Contents ABSTRACT ..................................................................................................................... ii ACKNOWLEDGEMENTS ............................................................................................ iv LIST OF TABLES .......................................................................................................... vii LIST OF FIGURES ........................................................................................................ viii ATTRIBUTIONS ............................................................................................................ x Chapter 1. Introduction........................................................................................................................ 1 Research Objectives ........................................................................................................... 6 Literature Cited .................................................................................................................. 7 Chapter 2. Small fluctuations in the recovery of fusaria across consecutive sampling intervals with unmanned aircraft 100 m above ground level....................... 13 Abstract .............................................................................................................................. 14 Introduction ........................................................................................................................ 15 Materials and Methods ....................................................................................................... 16 Results…………………..................................................................................................... 18 Discussion .......................................................................................................................... 21 Acknowledgements ............................................................................................................ 24 Literature Cited .................................................................................................................. 25 Chapter 3. Diverse assemblages of fusaria collected with unmanned aircraft 100 m above ground level suggest contribution of multiple sources across broad geographical regions.......................................................................................................... 36 Abstract .............................................................................................................................. 37 Introduction ........................................................................................................................ 39 Materials and Methods ....................................................................................................... 41 Results ................................................................................................................................ 47 Discussion .......................................................................................................................... 51 Acknowledgements ............................................................................................................ 56 Literature Cited .................................................................................................................
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